Abstract
Nanos and pumilio bind each other to regulate translation of specific mRNAs in germ cells of model organisms, such as D. melanogaster or C. elegans. Recently described human homologues NANOS1 and PUMILIO2 form a complex similar to their ancestors. This study was aimed to identify the proteins interacting with NANOS1-PUMILIO2 complex in the human spermatogenic cells. Here, using the yeast two-hybrid system we found that NANOS1 and PUMILIO2 proteins interact with RNA DEAD-box helicase GEMIN3, a microRNA biogenesis factor. Moreover, GEMIN3 coimmunoprecipitates with NANOS1 and PUMILIO2 in transfected mammalian cells. By double immunofluorescence staining, we observed that complexes built of NANOS1, PUMILIO2 and GEMIN3 are located within cytoplasmic region of germ cells. These proteins condense to form a compact aggregate in the round spermatids of the human and mouse germ cells. This aggregate was reminiscent of the chromatoid body (CB), a perinuclear structure present in the mammalian male germ line. This structure is considered evolutionary remnant of germ plasm, a hallmark structure of germ cells in lower metazoan. Using a CB marker VASA protein, we demonstrated that CBs are present in the human round spermatids, as they are in the mouse. Moreover, NANOS1, PUMILIO2 and GEMIN3 colocalize with VASA protein. We demonstrated for the first time that a mammalian Nanos-Pumilio complex functions within CB, a center of RNA storing and processing, involving microRNAs. NANOS1-PUMILIO2 complex, together with GEMIN3 and small noncoding RNAs, possibly regulate mRNA translation within CB of the human germ cells.
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Acknowledgments
We thank Dr. Robert P Erickson and the members of Jaruzelska lab for comments on the manuscript, Aleksander Ratajczyk and Marcin Kujawa from laboratory of confocal microscopy at Adam Mickiewicz University for assistance while analyzing fluoroimmunohistochemical reactions, Dr. Krzysztof Deregowski for providing us with prenatal testis tissue sections and Dr. Friedrich A Grässer with anti-GEMIN3 antibodies. This work was supported by Polish Ministry of Science and Higher Education (grant numbers N401129 32/2581 and N N401280833 to J.J., 6PO5E 001 21 and 3PO5E 013 25 to K.K.).
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Ginter-Matuszewska, B., Kusz, K., Spik, A. et al. NANOS1 and PUMILIO2 bind microRNA biogenesis factor GEMIN3, within chromatoid body in human germ cells. Histochem Cell Biol 136, 279–287 (2011). https://doi.org/10.1007/s00418-011-0842-y
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DOI: https://doi.org/10.1007/s00418-011-0842-y